title: Stimulation of fibroblasts and neuroblasts on a biomimetic extracellular matrix consisting of tandem repeats of the elastic VGVPG domain and RGD motif.
abstract: Elastin-like proteins (ELPs) modeled after tropoelastin are favored in the development of biomimetic matrices due to their biocompatibility and the possibility to precisely control their environmental responsiveness, mechanical properties, and fate within the cells by recombinant DNA technology-mediated design at the gene level. However, a basic prerequisite in the use of ELPs as cell culture matrices is the presence of a biofunctionality that can induce adhesion-mediated signaling pathways. To activate fibronectin-integrin signaling events from a cell-matrix interface and direct cell survival and proliferation, we biosynthesized a modular ELP, represented as TGPG[VGRGD(VGVPG)₆]₂₀ WPC, consisting of alternating elastic (VGVPG)₆structural domains and cell-binding VGRGD motifs that are intended to emulate various aspects of extracellular matrix proteins. The inverse transition curves of [VGRGD(VGVPG)₆]₂₀ and (VGVPG)₁₄₀ overlapped with each other, indicating that one VGRGD sequence fused with six elastic pentapeptides did not disturb the thermal sensitivity of [VGRGD(VGVPG)₆]₂₀. The cell adhesion activity of [VGRGD(VGVPG)₆]₂₀ toward HEK293 fibroblasts and N2A neuroblasts was similar to that of native fibronectin. Upon contact with [VGRGD(VGVPG)₆]₂₀, the fibroblasts exhibited a flattened polygonal morphology, and the neuroblasts synthesized new DNA and proliferated. On the basis of these physiological changes, we concluded that RGD-functionalized ELP triggers the activation of signaling cascades within cells and can be used as an elastin-like matrix for mammalian cell culture.
authors.affiliation: Laboratory of Biochemistry and Cellular Engineering, Daegu Gyeongbuk Institute of Science and Technology, Daegu 704-230, Republic of Korea. wbjeon@dgist.ac.kr
authors.forename: Won Bae
authors.initials: WB
authors.lastname: Jeon
authors.forename: Bo Hyung
authors.initials: BH
authors.lastname: Park
authors.forename: Junjun
authors.initials: J
authors.lastname: Wei
authors.forename: Rang-Woon
authors.initials: RW
authors.lastname: Park
mesh_terms: D032701:Biomimetics; D002448:Cell Adhesion; D002460:Cell Line; D049109:Cell Proliferation; D002470:Cell Survival; D003198:Computer Simulation; D004548:Elasticity; D005109:Extracellular Matrix / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism; D006801:Humans; D009474:Neurons / Q000378:metabolism; D009842:Oligopeptides / Q000737:chemistry; D017434:Protein Structure, Tertiary; D015398:Signal Transduction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D013696:Temperature
chemical_list: D005353:Fibronectins; D009842:Oligopeptides; C047981:arginyl-glycyl-aspartic acid